Cryopreservation of winter-dormant apple: III Bud water status and survival after cooling to -30°c and during recovery from cryopreservation

Abstract In a continuing study to improve the efficiency of dormant bud cryopreservation for tissues hardened in maritime climates, the water status of dormant buds was monitored between -4°C and recovery from liquid nitrogen (LN). Measurement of water content, simple thermal analysis and differential scanning calorimetry were employed. Buds did not lose water during cooling to, or holding at -30°C indicating that cryodehydration and/or other adaptive responses contributed during this essential step. A bud exotherm that was an artefact of warming was detected due to necessary handling at -4°C before cooling to -30°C. There were no significant differences between cultivars with respect to water status at -30°C or immediately upon rewarming from LN despite significant differences in post-LN survival. Buds rehydrated in 5 days, but up to 14 days may be needed for recovery for some cultivars. In some instances buds could be grafted without rehydration, taking up water across the early graft union

Molecular evidence of Cacao swollen shoot virus acquisition and retention by Planococcus citri (Risso) and Pseudococcus longispinus (Targioni-Tozzetti) and Pseudococcus viburni (Signoret) Mealybugs (Hemiptera: Pseudococcidae)

Cacao swollen shoot virus (CSSV) remains a significant viral pathogen endemic in West African cacao-growing countries. Eradication of affected trees (complete removal) and resistance breeding in existing cacao (Theobroma cacao L.) germplasm have not yielded the expected results in terms of controlling the spread of CSSV. CSSV is reported to be vectored by at least 16 species of mealybugs (Hemiptera: Pseudococcidae), but there have been no molecular evidences to validate these reports. Inconsistent reports exit on CSSV transmission which were based on visual assessments of CSSV infection which are highly subjective, especially on asymptomatic cacao. The validation of the transmission efficiency of CSSV by mealybugs are arguable. In this study, the presence of CSSV in three mealybug species, Planococcus citri (Risso), Pseudococcus longispinus (Targioni Tozzetti) and Pseudococcus viburni (Signoret), was monitored via PCR after a 72-h virus acquisition access period (AAP) on symptomatic T. cacao (var. Amelonado) seedlings ‘infected’ with New Juaben, a severe strain of CSSV. Solanum tuberosum sprouts were then fed to the mealybugs over a sixday period. Sequenced PCR products of the destructively sampled and CSSV-screened individual mealybug species provided a molecular evidence of CSSV acquisition. qPCR results gave another evidence that CSSV retention is highly variable between and within mealybug species. This is the first molecular evidence highlighting a non-West African mealybug species, Ps. viburni, as a capable CSSV vector. Apparently, these results could validate the efficacy of non-malvaceous species as barrier crops and in screening for resistance to CSSV vectors in T. cacao

Cryopreservation of winter-dormant apple buds: II - tissue water status after desiccation at -4°c and before further cooling

Abstract The established protocol for the cryopreservation of winter-dormant Malus buds requires that stem explants, containing a single, dormant bud are desiccated at -4°C, for up to 14 days, to reduce their water content to 25-30% of fresh weight. Using three apple cultivars, with known differences in response to cryopreservation, the pattern of evaporative water loss has been characterised, including early freezing events in the bud and cortical tissues that allow further desiccation by water migration to extracellular ice. There were no significant differences between cultivars in this respect or in the proportions of tissue water lost during the desiccation process. Differential Scanning Calorimetry (to -90°C) of intact buds indicated that bud tissues of the cultivar with the poorest response to cryopreservation had the highest residual water content at the end of the desiccation process and froze at the highest temperature Keywords: Malus, cryopreservation, dormant bud, dehydratio

Increase of the mean inner Coulomb potential in Au clusters induced by surface tension and its implication for electron scattering

Electron holography in a transmission electron microscope was applied to measure the phase shift induced by Au clusters as a function of the cluster size. Large phase shifts Df observed for small Au clusters cannot be described by the well-known equation Df=C_E V_0 t (C_E: interaction constant, V_0: mean inner Coulomb potential (MIP) of bulk gold, t: cluster thickness). The rapid increase of the Au MIP with decreasing cluster size derived from Df, can be explained by the compressive strain of surface atoms in the cluster

Sexual health clinic attendance and non-attendance in Britain: findings from the third National Survey of Sexual Attitudes and Lifestyles (Natsal-3)

Objectives In Britain, sexual health clinics (SHCs) are the most common location for STI diagnosis but many people with STI risk behaviours do not attend. We estimate prevalence of SHC attendance and how this varies by sociodemographic and behavioural factors (including unsafe sex) and describe hypothetical service preferences for those reporting unsafe sex.Methods Complex survey analyses of data from Britain’s third National Survey of Sexual Attitudes and Lifestyles, a probability survey of 15 162 people aged 16–74 years, undertaken 2010–2012.Results Overall, recent attendance (past year) was highest among those aged 16–24 years (16.6% men, 22.4% women), decreasing with age (<1.5% among those 45–74 years). Approximately 15% of sexually-active 16–74 year olds (n=1002 men; n=1253 women) reported ‘unsafe sex’ (condomless first sex with a new partner and/or >=2 partners and no condom use, past year); >75% of these had not attended a SHC (past year). However, of non-attenders aged 16–44 years, 18.7% of men and 39.0% of women reported chlamydia testing (past year) with testing highest in women aged <25 years. Of those aged 16–44 years reporting unsafe sex, the majority who reported previous SHC attendance would seek STI care there, whereas the majority who had not would use general practice.Conclusion While most reporting unsafe sex had not attended a SHC, many, particularly younger women, had tested for chlamydia suggesting engagement with sexual health services more broadly. Effective, diverse service provision is needed to engage those at-risk and ensure that they can attend services appropriate to their needs

Mechanical Mitral Valve Replacement: A Multicenter Study of Outcomes with Use of 15- to 17-mm Prostheses

Background: The aim of this study was to evaluate early and mid-term outcomes (mortality and prosthetic valve reintervention) after mitral valve replacement with 15- to 17-mm mechanical pr

Elimination of onchocerciasis in Ecuador: findings of post-treatment surveillance.

BACKGROUND: The Esmeraldas focus of onchocerciasis in Ecuador expanded geographically during the 1980s and was associated with severe ocular and skin disease. Mass drug administration (MDA) with ivermectin started in 1991, initially once but later twice a year, in the principle endemic focus followed by all satellite foci. Treatment was stopped in 2009 when entomological assessments determined that transmission of Onchocerca volvulus had been interrupted. METHODS: Three years after the cessation of ivermectin treatment in 2012, as defined by the WHO guidelines for onchocerciasis elimination, blackfly collections were done in four sentinel sites in former hyperendemic areas. The presence of infective larvae in local vectors, Simulium exiguum and Simulum quadrivittatum, was assessed by detection of O. volvulus DNA by PCR. Additional flies captured in four extra-sentinel sites located in former hyper- and mesoendemic dispersed isolated areas were also assessed. RESULTS: The results from 68,310 captured blackflies, 40,114 from four sentinel villages in the previously hyperendemic areas (Corriente Grande, El Tigre, San Miguel on Río Cayapas and Naranjal on Río Canandé) and 28,197 from extra-sentinel locations, were all negative for the presence of O. volvulus. These extra-sentinel sites (Hualpí on Río Hoja Blanca, Capulí on Río Onzole, La Ceiba on Río Tululví and Medianía on Río Verde) were included to provide additional evidence of the impact of MDA on the transmission of O. volvulus in isolated endemic areas. CONCLUSIONS: Our data indicate that transmission of O. volvulus has been stopped in all endemic areas in Ecuador, including all satellite foci outside the main focus. These findings indicate that a strategy of ivermectin distribution twice a year to over 85% of the treatment-eligible population was effective in eliminating the infection from Ecuador in a focus with a highly competent primary vector, S. exiguum, and where the infection rates were equal to or greater than observed in many onchocerciasis foci in Africa

Density-Dependent Mortality of the Human Host in Onchocerciasis: Relationships between Microfilarial Load and Excess Mortality

Human onchocerciasis (River Blindness) is a parasitic disease leading to visual impairment including blindness. Blindness may lead to premature death, but infection with the parasite itself (Onchocerca volvulus) may also cause excess mortality in sighted individuals. The excess risk of mortality may not be directly (linearly) proportional to the intensity of infection (a measure of how many parasites an individual harbours). We analyze cohort data from the Onchocerciasis Control Programme in West Africa, collected between 1974 and 2001, by fitting a suite of quantitative models (including a ‘null’ model of no relationship between infection intensity and mortality, a (log-) linear function, and two plateauing curves), and choosing the one that is the most statistically adequate. The risk of human mortality initially increases with parasite density but saturates at high densities (following an S-shape curve), and such risk is greater in younger individuals for a given infection intensity. Our results have important repercussions for programmes aiming to control onchocerciasis (in terms of how the benefits of the programme are calculated), for measuring the burden of disease and mortality caused by the infection, and for a better understanding of the processes that govern the density of parasite populations among human hosts